The Why and How of: Reagent / QC Lot Roll-over Validation

The Why and How of:
Reagent / QC Lot Roll-over Validation
LabLink Interlab Reports
Sandy Gardner B.S., M.T. (ASCP)
Mary Ann Kozy
Carol Shearer B.S., CLS (NCA), M.T. (ASCP)
Dade Behring Hemostasis Application Consultants
Hemostasis Application Consultants
Carol Shearer
Region I West
Sandy Gardner
Region II Central
Mary Ann Kozy
Region III East
Hemostasis Application Consultants
Region I – Carol Shearer
Region II – Sandy Gardner
Region III – Mary Ann Kozy
Hemostasis Application Consultant
 Scope
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Field support for Clinical Application Specialist & Field Service Reps
Assay application and software
Training…customers and Dade Behring personnel
Seminar Presentations
Instrument Evaluations
IDN Instrument Implementation
Troubleshooting Escalation
 Support
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40
• 146
• 4200
Sales Regions
CAS’s
FSR’s
Hemostasis analyzers
 Special Projects
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Marketing
Second Level Support
America’s-Product Education Training
Sales
Global projects
Clinical Application Specialist (CAS)
• Working directly with the lab team on-site,
responsibilities include:
• On-site instrument customization and validation
• On-site training of primary operator to perform basic
operation, calibration, maintenance and
troubleshooting
• Normal Range and Precision studies, Method
Correlations, Heparin Therapeutic Range, D-Dimer
ROC curves
• Statistical analyses of data for the lab and summary
report
• On-going training for laboratory as needed
The Why and How of:
Reagent / QC Lot Roll-over Validation
Sandy Gardner B.S., M.T. (ASCP)
Region II
Dade Behring Hemostasis Application Consultant
Lot Roll-over…Regulatory Agency Requirements
 CLIA ’88
• 493.1255
• 493.1269
Calibration
PT MNPT for INR calculation
 CAP
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HEM.23360
HEM.23430
HEM.23453
HEM.23476
HEM.23500
HEM.23575
• HEM.24575
• HEM.38006
• HEM.38008
GeoMean MNPT PT sec
Check INR calculation
aPTT Therapeutic Range
aPTT Therapeutic Range establishment
Reference Intervals
Recommendation for anticoagulant therapeutic
range
New reagent lots vs. old reagent lots
New lot calibration
Quality Control acceptable range
Lot Roll-over…References
 CLSI
• H47-A 1996
• EP5-A 1998
• EP15-A 2002
• EP18-A 2002
One Stage Prothrombin Time (PT)
Evaluation of precision performance of
clinical chemistry devices
User demonstration of performance for
precision and accuracy
Quality Management for unit-use testing
Calibration
 Calibrated assays
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Fibrinogen
Factors
D-Dimer
ATIII
Protein C and Protein S
vWF
alpha-2 Antiplasmin
Plasminogen
Heparin Xa
 Frequency
• Reagent lot number change
• Verification every 6 months
Calibration
 Curve
• Minimum 3 points
 Zero
 Mid point value
 Maximum value near upper limit of assay
Reference Interval...PT and aPTT
 Reference Interval Verification
(Normal Range Study)
• Minimum 20 samples
• Represent laboratory’s healthy population guidelines
 No hospital/clinic patients
 Questionnaire
 10 males / 10 females
 a priori criteria with exclusion before testing
 a posteriori criteria with exclusion after testing
• Exclude outliers and replace with additional
sample
• Acceptability of 95 % range
 +/- 10 % within range
 > 10% outside of range another 20 samples tested
Reference Interval...PT and aPTT
 Sample Criteria
• 3.2 % sodium citrate
• Process per CLSI/CAP Guidelines
 PT
 aPTT
uncentrifuged/centrifuged unopened 18-24oC 24 hours
uncentrifuged/centrifuged unopened 2-4oC or
18-24oC 4 hours
 Platelet poor plasma @ < 10,000/uL
• Frozen specimens CLSI Guidelines
 -20oC for 2 weeks
 -70oC for 6 months
 Thaw @ 37oC and test immediately or max 2 hours @ 4oC
PT MNPT for INR Calculation
 Geometric Mean of Reference Interval
• More appropriate estimate of the average value
 Geometric Mean Calculation
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GM=antilog[(log(X1)+log(X2)+log(X3)+…log(Xn)/n]
Microsoft Excel
EP Evaluator version 7
www.graftacs.com/geomean.php3
 INR Calculation
• INR =
PT sec Patient
GeoMNPT sec
ISI
PT MNPT for INR Calculation
 Verification of INR Calculation
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Yearly
Change in lot, reagent, instrument
Establishment of new PT reference range
Change in INR calculation
 Check INR calculated by analyzer or LIS
• Check @ 2.0 and 3.0 INR
• Manual calculation
• Microsoft Excel calculation
aPTT Unfractionated Heparin Therapeutic Range
 aPTT vs. Heparin Xa assay
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30 – 40 patients receiving only UFH
Samples should cover the assay range
INR <1.3 to assure no concomitant Coumadin therapy
5 – 10 normal samples
aPTT processed and assayed per CLSI Guidelines
Heparin Xa assayed fresh or frozen
 Double centrifuge sample
 Platelet poor plasma <10,000/uL
 CLSI storage guidelines
 Calculation of Therapeutic Range
• Plot Heparin Xa IU/mL (X) vs. aPTT seconds (Y)
• Heparin Xa Therapeutic Range = 0.3 – 0.7 IU/mL
• Calculate aPTT Range from Regression Line
 Lower aPTT value = Slope (0.3) + Intercept
 Upper aPTT value = Slope (0.7) + Intercept
aPTT Unfractionated Heparin Therapeutic Range
200
180
Therapeutic Range
160
a
P 140
T
T
120
Hep Xa
0.3
0.7
s
100
e
c
o 80
n
d 60
s
40
N=
Corr Coef (R)
Slope
Intercept
20
0
0.00
0.20
0.40
0.60
0.80
Heparin U/mL
1.00
1.20
1.40
aPTT
65
95
77
0.67
75.7
41.9
Things to Consider
 INR > 1.3 should not be used
 Evaluate “unusual” results
• Elevated aPTT with 0.0 Heparin Xa
• Low aPTT with elevated Heparin Xa
• Must justify deletion of data point
 Consider
• Sample processing
• Direct Thrombin Inhibitors
• Other drugs
 Avoid
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Insufficient samples
aPTT values above reportable range
Heparin Xa values above calibration curve
More than 2 samples per patient unless dose change
aPTT Therapeutic Range Validation
 Establish initial reagent lot using Heparin Xa vs. aPTT
 Subsequent reagent lot validation based on Method Correlation
 20 - 30 samples (statistically significant) covering range of assay
 Plot current lot on X axis & new lot on Y axis
 Evaluate regression data
 Record difference between the means of old & new
 Year 3, 4, etc. determine cumulative mean difference
 Acceptable difference of mean or cum mean = < 7 seconds
Validation of Heparin Sensitivity
with Existing, Validated aPTT Reagent
Year
Mean Current
Mean New
New – Current
Cum Sum
Accept
2001
78.6
73.9
-4.7
- 4.7
yes
2002a
48.3
41.7
-6.6
-11.3
no
2002b
47.6
53.6
+6.0
+ 1.3
yes
2003
71.9
72.3
+0.4
+ 1.7
yes
2004a
62.0
71.2
+9.2
+10.9
no
2004b
62.8
60.3
-2.5
- 0.8
yes
A difference of > 7 sec between Means or change in Cum Sum requires
evaluation and action. 0 – 5 sec preferred.
ACTION for > 7 second difference:
1.
Evaluate second lot
2.
Reestablish Therapeutic Range
Method Verification (Correlation)
 Patient sample comparisons recommended as
good laboratory science
 Normal samples “rarely” change lot to lot
 Abnormal samples detect differences in reagent
sensitivity
 Allows comparison of assay data range to detect
differences in patient result recovery
Method Verification Study
 40 patient samples
 20 normal + 20 abnormal
 Values cover range of assay normal to abnormal
 Assay on current and new lots within 1 hour optimally
 Calculate data with regression analysis
 Lot to Lot Consistency
• Corr Coef (R)
• Slope
• Intercept
0.90 – 0.95
0.96 – 0.98
0.99 – 1.00
1.00 +/- 0.05
0.00 +/- 95%
acceptable
good
excellent
Quality Control Ranges
 Establish QC ranges for new lot numbers
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New lot reagent + new lot QC material
Current lot reagents + new lot QC material
Unassayed controls
Assayed controls…recommend establish lab range
 Establish Range
• Minimum 20 measurements
 20 working days
 10 working days / 2 runs per day
 Outliers…discard and replace
• Multiple vials, multiple techs, multiple days builds
system variability into data
 QC Data Significant Figure
• One more significant digit than reportable value
• Provides better mean and SD for data calculation
Quality Control
 Data Calculation
• Mean……average of data
• SD……….accounts for random error in system
• % CV……variation around the mean
 Coagulation Quality Control
• Every 8 hours of testing
• 2 levels minimum
 Normal level
 Abnormal level
Tips & Tricks for Lot Roll-over
 Communicate with QAP Coordinators
• Establish volumes
• Establish ship dates
• Monitor usage
 Automated analyzers
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Simultaneous assay of current and new lot numbers
PT vs. New PT
aPTT vs. New aPTT
Current QC with current reagent lots
New QC with new reagent lots or current lots
 Dade Behring analyzers…refer to Lot Roll-over procedures
• Contact local CAS or TAC
The Why and How of:
LabLink Interlab Reports
Carol Shearer B.S.,CLS (NCA), M.T.(ASCP)
Region I
Dade Behring Hemostasis Application Consultant
Interlab Program…What Is It ?
 Comparison of lab data to group data
 Gives reference point to assure system is in control
 Additional Tool
• Assess stability of test system
• Aid in setting QC limits and rules
• Aid in troubleshooting
Interlaboratory Survey Group
 QC lot number specific
 Analyte specific
 Reagent type not lot specific
 Same instrument + all instruments
 Data submitted monthly
LabLink® On-line Set-up
 www.dadebehring.com
 Register
 Configure
 Tools
• LabLink® On-line Quick Instructions
• QAP Coordinator 1-800-242-DADE opt 4 opt 2
Interlaboratory Report Data Entry
 Submit monthly
• N (number of data points)
• Mean
• SD
Lab Statistics Report
Individual Lab Report
Individual Lab Report Data
Mean
MTD & LTD
SD
CV
# points
Individual Lab Report Data
Test System Peer
SDI
CVI
SDI = Lab mean-Group Mean
Group SD
CVI = Lab CV / Group CV
Individual Lab Report Data
Total Error = % bias + (2 x CV)
% bias = (Lab mean – Group mean)x100 / Group mean
Total Error
A
Lab + 2SD
Lab Mean
Lab – 2SD
Peer Mean
C
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Lab’s
Imprecision
B
x
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Lab’s
Total
Error
Lab’s
Bias
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Individual Lab Report Total Error Graph
Monthly Summary Report
Exception Notes Report
Condensed Interlab Report
Value of Interlaboratory Program
 Allows comparison to many other labs
 Aids in setting QC limits
 Aids in assessing accuracy and precision
 Aids in troubleshooting
 Assists in evaluating lot to lot variation
Web Sites
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www.dadebehring.com
www.cap.org
http://wwwn.cdc.gov/clia/regs/subpart_k
www.dgrhoads.com
www.westgard.com
www.clsi.org
Questions & Answers
 To ask a question in the webcast window:
– Type your question in the Question Box below the
presentation.
– Click Submit.
 To ask a question via the telephone:
– Dial (800) 661-2563 .
– Press *1 on your touch-tone keypad.
Thank You !
For assistance with Dade Behring Hemostasis products contact the
local Dade Behring Sales Representative, CAS or TAC.
For LabLink® assistance contact the Dade Behring QAP Coordinator
1-800-242-DADE
References
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College of American Pathology Hematology-Coagulation Checklist, revised 10-31-06
CLIA ’88, Subpart K, Quality Systems for Nonwaived Testing
CLSI, H47-A, One-Stage Prothrombin Time (PT) Test and Activated Partial Thromboplastin
Time (APTT) Test, June 1996
CLSI, EP5-A, Evaluation of Precision Performance of Quantitative Measurement Methods,
August 2004
CLSI, EP15-A2, User Verification of Performance for Precision and Trueness, April 2006
CLSI, EP18-A, Quality Management for Unit-Use Testing, 2002
CLSI, C28-A2, How to Define and Determine Reference Intervals in the Clinical Laboratory,
June 2000
CLSI, H21-A4, Collection, Transport, and Processing of Blood Specimens for Testing
Plasma-based Coagulation Assays, December 2003
Establishing APTT Values Corresponding to the Heparin Therapeutic Range, Dade Behring,
Hemostasis Technical Bulletin 01-2004, January 2004
CAP Today, How to Validate Heparin Sensitivity of aPTT, Dr. John Olson, October 2004
Basic QC Practices 2nd Edition, Training in Statistical Quality Control for Healthcare
Laboratories, James O. Westgard, Ph.D., 2002
EP Evaluator version 7, David Rhoads, Interpretation Guidelines, 2005
Dade Behring Control Plasma N and Control Plasma P package insert, January 2005
Dade Behring LabLink® Quaity Assurance Program